PMID- 28501500
OWN - NLM
STAT- MEDLINE
DCOM- 20180406
LR  - 20191210
IS  - 1879-1166 (Electronic)
IS  - 0198-8859 (Linking)
VI  - 78
IP  - 7-8
DP  - 2017 Jul-Aug
TI  - It's about time: The development and validation of a rapid optimized single 
      antigen bead (ROB) assay protocol for LABScreen.
PG  - 489-499
LID - S0198-8859(17)30073-3 [pii]
LID - 10.1016/j.humimm.2017.05.001 [doi]
AB  - The LABScreen single antigen bead assay (SAB) is a method widely used for the 
      identification and monitoring of human leukocyte antigen (HLA) antibodies in 
      patients pre-and post-transplant. While accurate testing of patient samples is 
      key for optimal patient care, time can also be important, especially during 
      deceased donor workups or post-transplant assessments. Here we describe the 
      development and validation of the Rapid Optimized SAB (ROB) protocol, a modified 
      version of the One Lambda LABScreen SAB (OLSAB) procedure, which reduces assay 
      time from 85 to 25min (>70% reduction) without impacting assay quality or 
      sensitivity. Optimization steps included shortened centrifugation cycles and 
      reduced serum and secondary antibody incubation times in combination with 
      increased secondary antibody concentration. Linear regression analysis of 
      baseline median fluorescence intensity (MFI) values showed excellent correlation 
      between the ROB and OLSAB protocols (r(2)>0.98) for both class I and class II 
      antibodies in 58 sera tested in two HLA laboratories. Importantly, the ROB 
      protocol demonstrated a trend towards improved inter-laboratory MFI concordance 
      when compared to the OLSAB procedure (r(2)=0.9816 vs 0.9451), especially for HLA 
      antibody specificities in the 500-2000 MFI range (r(2)=0.7824 vs 0.6313). 
      Implementation of the ROB protocol will expedite HLA antibody testing and may 
      improve reproducibility of the SAB assay.
CI  - Copyright (c) 2017 American Society for Histocompatibility and Immunogenetics. 
      Published by Elsevier Inc. All rights reserved.
FAU - Liwski, Robert S
AU  - Liwski RS
AD  - Department of Pathology, Dalhousie University, Halifax, Nova Scotia B3H 1V8, 
      Canada. Electronic address: robert.liwski@nshealth.ca.
FAU - Greenshields, Anna L
AU  - Greenshields AL
AD  - Department of Pathology, Dalhousie University, Halifax, Nova Scotia B3H 1V8, 
      Canada.
FAU - Murphey, Cathi
AU  - Murphey C
AD  - Southwest Immunodiagnostics Inc., San Antonio, San Antonio, TX 78229, USA.
FAU - Bray, Robert A
AU  - Bray RA
AD  - Department of Pathology, Emory University Hospital, Atlanta, GA 30322, USA.
FAU - Gebel, Howard M
AU  - Gebel HM
AD  - Department of Pathology, Emory University Hospital, Atlanta, GA 30322, USA.
LA  - eng
PT  - Journal Article
PT  - Validation Study
DEP - 20170510
PL  - United States
TA  - Hum Immunol
JT  - Human immunology
JID - 8010936
RN  - 0 (Antibodies)
RN  - 0 (HLA Antigens)
SB  - IM
MH  - Antibodies/*blood
MH  - Graft Rejection/*diagnosis/immunology
MH  - HLA Antigens/immunology
MH  - Humans
MH  - *Immunoassay
MH  - Microspheres
MH  - Monitoring, Physiologic/*methods
MH  - Observer Variation
MH  - Reproducibility of Results
MH  - Sensitivity and Specificity
MH  - Time Factors
MH  - *Transplantation
OTO - NOTNLM
OT  - Assay optimization
OT  - HLA antibodies
OT  - Single antigen bead assay
OT  - Transplantation
EDAT- 2017/05/16 06:00
MHDA- 2018/04/07 06:00
CRDT- 2017/05/15 06:00
PHST- 2017/02/16 00:00 [received]
PHST- 2017/04/27 00:00 [revised]
PHST- 2017/05/05 00:00 [accepted]
PHST- 2017/05/16 06:00 [pubmed]
PHST- 2018/04/07 06:00 [medline]
PHST- 2017/05/15 06:00 [entrez]
AID - S0198-8859(17)30073-3 [pii]
AID - 10.1016/j.humimm.2017.05.001 [doi]
PST - ppublish
SO  - Hum Immunol. 2017 Jul-Aug;78(7-8):489-499. doi: 10.1016/j.humimm.2017.05.001. 
      Epub 2017 May 10.